Selection of reference genes for qRT-PCR analysis of gene expression in sea cucumber Apostichopus japonicus during aestivation

Ye Zhao; Muyan Chen; Tianming Wang; Lina Sun; Dongxue Xu; Hongsheng Yang

doi:10.1007/s00343-015-4004-2

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Selection of reference genes for qRT-PCR analysis of gene expression in sea cucumber Apostichopus japonicus during aestivation

Updated：2023-05-16

- Selection of reference genes for qRT-PCR analysis of gene expression in sea cucumber Apostichopus japonicus during aestivation
- Journal of Oceanology and Limnology Vol. 32, Issue 6, Pages: 1248-1256(2014)
- Affiliations：
  
  1. Institute of Oceanology Chinese Academy of Sciences
  2. University of Chinese Academy of Sciences
  3. Ocean University of China
  4. Zhejiang Ocean University
- Author bio：
- Funds：
- DOI：10.1007/s00343-015-4004-2
  CLC：
- Online First：18 July 2014，
  
  Published：2014
- Accepted：
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Zhao, Y., Chen, M., Wang, T. et al. Selection of reference genes for qRT-PCR analysis of gene expression in sea cucumber Apostichopus japonicus during aestivation., Chin. J. Ocean. Limnol. 32, 1248–1256 (2014). https://doi.org/10.1007/s00343-015-4004-2
DOI：

Zhao, Y., Chen, M., Wang, T. et al. Selection of reference genes for qRT-PCR analysis of gene expression in sea cucumber Apostichopus japonicus during aestivation., Chin. J. Ocean. Limnol. 32, 1248–1256 (2014). https://doi.org/10.1007/s00343-015-4004-2 DOI：

摘要

Abstract

Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) is a technique that is widely used for gene expression analysis

and its accuracy depends on the expression stability of the internal reference genes used as normalization factors. However

many applications of qRT-PCR used housekeeping genes as internal controls without validation. In this study

the expression stability of eight candidate reference genes in three tissues (intestine

respiratory tree

and muscle) of the sea cucumber Apostichopus japonicus was assessed during normal growth and aestivation using the geNorm

NormFinder

delta CT

and RefFinder algorithms. The results indicate that the reference genes exhibited significantly different expression patterns among the three tissues during aestivation. In general

the β-tubulin (TUBB) gene was relatively stable in the intestine and respiratory tree tissues. The optimal reference gene combination for intestine was 40S ribosomal protein S18 (RPS18)

TUBB

and NADH dehydrogenase (NADH); for respiratory tree

it was β-actin (ACTB)

TUBB

and succinate dehydrogenase cytochrome B small subunit (SDHC); and for muscle it was α-tubulin (TUBA) and NADH dehydrogenase [ubiquinone] 1 α subcomplex subunit 13 (NDUFA13). These combinations of internal control genes should be considered for use in further studies of gene expression in A. japonicus during aestivation.

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references

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